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Fig. 2. UV-exposed cells did not reveal increased phosphate incorporation in the
EGF receptor. In vivo labelling of cells were performed by preincubation of
cells with phosphate-free medium (MEM) for 2 hours, followed by incubation
with [32P] orthophosphate in DMEM for 2 hours. Cells were then left
untreated or exposed to UV or EGF, and incubated for the indicated time
intervals. Cell lysates were subject to immunoprecipitation with an
anti-EGFR-protein G-sepharose conjugate. The EGFR immunoprecipitates were
separated by SDS-PAGE, and identified by (A) autoradiography and (B) western
immunoblotting with anti-EGFR. Cells stimulated with 5 nM EGF for 5 minutes
were used as positive controls. C and E denote control and EGF-stimulated
cells, respectively. The molecular weight markers presented at the left
indicate 220 kDa.